Part:BBa_K2570011

dsrA+lacI+PlacI+hns+ProU+GFP

In our project, we combined temperature promoter dsrABBa_K2570015 and salt-concentration promoter proUBBa_K2467001 as a logical gate controlled by salt-concentration and temperature to realize the aim of producing antibacterial, PLA. dsrABBa_K2570015 is a temperature control promoter. The temperature range of dsrA is from 25 degree to 42 degree. With temperature increase, the promote efficiency is getting higher. [1]The salt-concentration promoter proUBBa_K2467001 provides sensitivity to the salt concentration to human skin. proU is a promoter existed in E.coli, induced by low salt concentration. And the H-NSBBa_K2570016 is a global-regulated protein which could bind to genome and specifically inhibit proU.[2]

In our design, our engineering bacteria usually put in a lotion container and store at room temperature. When temperature stays below 37 degree, the temperature promoter will not work.

Fig.1 our circuits in room temperature.

The function of PlacI BBa_R0011 has become a constitutive promoter. The global regulate protein H-NS will be expressed to inhibit proU. Then the logical gate will be “off” state.

On the other hand, when we use the lotion on the skin, temperature reached 37 degree, the repressor protein LacI BBa_C0012 will be expressed to inhibit PlacI, the H-NS won’t be revealed. Then, once people sweat, the Na+ and Cl- will induce proU, the logical gate became “on” state.

Fig.1 our circuits on people skin.

Firstly we use GFP to characterize the function of our circuit. And after finishing the construction of the whole circuits, we set 3 gradient of salt concentration, 0M, 0.05M, 0.3M, and 3 gradient of temperature, 25℃, 32.5℃ and 37℃, detected the fluorescence level and draw the line of different culture conditions.

Fig.1 fluorescence level at different salt concentration and temperature.

Fig.2 the line of fluorescence level.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1587
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 3324
  Illegal SapI.rc site found at 154
  Illegal SapI.rc site found at 1973